The ECI2 Knockout Raji Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for targeted disruption of the ECI2 gene in the Raji B lymphocyte line. This product offers a heterogeneous pool of cells with ECI2 loss-of-function, enabling investigation of mitochondrial fatty acid ??-oxidation without the selective pressures of clonal isolation. The polyclonal format mirrors natural genetic variability while abolishing ECI2 function, providing a robust model for studying metabolic dependencies in lymphocyte biology and lymphomagenesis.
The Raji host cell line is an Epstein-Barr virus (EBV)-positive lymphoblastoid line derived from a Burkitt lymphoma patient. Retaining key B-cell characteristics, these cells are a standard model for immunological research, B-cell signaling, and apoptosis. Their continuous proliferation in culture and active oxidative metabolism render them particularly suitable for examining lipid catabolism and mitochondrial function, thereby facilitating high-throughput metabolic screening and functional genomics studies in a lymphoma-relevant context.
ECI2 encodes mitochondrial enoyl-CoA delta isomerase 2, a critical auxiliary enzyme in the ??-oxidation of unsaturated fatty acids. It catalyzes the isomerization of 3-cis and 3-trans enoyl-CoA intermediates to 2-trans enoyl-CoA, thereby channeling unsaturated fatty acid derivatives into the core ??-oxidation cycle. This process generates acetyl-CoA, NADH, and FADH2, which fuel the electron transport chain and ATP synthesis. ECI2 expression is positively regulated by PPAR??, PPAR??, ERR??, and the coactivator PGC-1??, linking its activity to nuclear receptor-mediated lipid metabolic programs. Within the ??-oxidation multienzyme complex, ECI2 functionally interacts with ACADVL, ACADM, ECHS1, HADHA, and HADHB. The broader pathway includes CPT1A for mitochondrial fatty acid import and ACOX1 in peroxisomal oxidation, illustrating a tightly coordinated system for lipid catabolism essential for cellular energy homeostasis.
In Raji B lymphocytes, ECI2-dependent isomerization is pivotal for complete unsaturated fatty acid degradation, sustaining lipid homeostasis and ATP production vital for proliferation and survival. Given the metabolic reprogramming typical of Burkitt lymphoma, ECI2 disruption may reveal exploitable metabolic vulnerabilities by causing incomplete oxidation, intermediate accumulation, and bioenergetic stress, thereby sensitizing lymphoma cells to metabolic insults and informing therapeutic strategies targeting fatty acid utilization.
Research applications include using these polyclonal knockout cells in CRISPR knockout screens to identify metabolic dependencies, mechanistic dissection of fatty acid oxidation in B-cell function, and preclinical evaluation of ECI2 as a cancer metabolic target. Typical assays encompass Western blotting and RT-qPCR for knockout validation, Seahorse metabolic flux analysis, fatty acid oxidation assays, cell viability and clonogenic studies, and flow cytometry for metabolic or apoptotic markers. For further information and support, please contact Ascent Research.
